羊水细胞作为组织工程种子细胞可能性的探讨  

Amniotic fluid: a novel source for tissue engineering

作　　者：李现铎[1] 耿红全[2] 潘骏[3] 谢华[2] 陈方[1] 

机构地区：[1]上海第二医科大学新华医院泌尿外科,200092 [2]上海第二医科大学新华医院小儿泌尿外科,200092 [3]上海儿科医学研究所

出　　处：《中华医学杂志》2005年第7期464-467,共4页National Medical Journal of China

基　　金：国家自然科学基金资助项目(30271351)

摘　　要：目的探讨羊水细胞作为一种新的组织工程种子细胞的可能性.方法通过羊膜腔穿刺取得人孕16～23周的羊水,分离细胞,在添加10%胎牛血清(FBS)的F10培养液中,体外培养传代,观察其生物学特性.免疫细胞化学方法检测细胞的标准中间丝蛋白,分析细胞性质.收集第4代细胞,种植到PGA生物支架上,通过倒置相差显微镜和扫描电镜观察细胞的黏附、生长情况.结果羊水细胞在体外培养体系中增殖迅速.免疫细胞化学显示波纹蛋白、α-平滑肌肌动蛋白、广谱角蛋白阳性;肌间线蛋白阴性,与间质细胞类似.羊水细胞能紧密的黏附在PGA生物支架上,呈汇合性生长,未发现细胞死亡的迹象.结论羊水细胞增殖能力强,表现间质细胞的特性,和生物支架有良好的相容性,具有作为组织工程种子细胞所需的条件.Objective To investigate the possibility of using amniotic fluid cells as seed cells for tissue engineering. Methods Amniotic fluid was obtained by ultrasound-guided amniocentesis performed on pregnant women with a gestational age ranging from 16~th ~23~rd weeks. The cells isolated from the amniotic fluid were cultured in F10 culture fluid with 10% FBS. Immunocytochemistry was used to examine the standard intermediate filaments. After 3 passages of subculture, the cells were harvested and seeded onto PGA polymer scaffold. The cellular morphology, structure and adhesion with scaffold were evaluated by contrast microscope and scanning electronic microscope. Results The amniocytes expanded rapidly in culture media. Immunocytochemistry revealed positive signals for vimentin, smooth muscle action (SMA), and pan cytokeratin, and negative signals for desmin. Amniocytes-polymer complex analysis showed confluent cells firmly attached to the scaffold, with no evidence of cell death. Conclusion The expansion potential of amniotic fluid cells is active. They express the characteristics of mesenchymal cells. The cells on PGA polymer scaffold can grow rapidly and maintain its morphological property. So the amniotic fluid may be a practical cell source for tissue engineering.

关 键 词：羊水细胞 种子细胞 组织工程 支架 间质细胞 体外培养 黏附 生长情况 种植 传代 

分 类 号：R318.08[医药卫生—生物医学工程]